Additive effect of the probiotics Lactobacillus exopolysaccharides and the Satureja calamintha extracts on enteropathogenic Escherichia coli adhesion

Abstract This study assessed the inhibitory potential of the probiotics Lactobacillus (LB) exopolysaccharides (EPS) with or without extracts of Satureja calamintha on enteropathogenic Escherichia coli (EPEc) responsible for gastroenteritis. Methanolic and hydromethanolic extracts were prepared by cold maceration and subjected to phytochemical screening. The compounds of the extracts were determined with the colorimetric assays and identified using high-performance liquid chromatography coupled with diode array detector (HPLC-DAD). Antioxidant activities of the extracts were also evaluated by using 2,2-diphenyl-1-picrylhydrazil (DPPH) radical scavenging. Antibacterial effect on EPEc was evaluated by using both agar disc diffusion and microdilution methods. The in vitro test of auto-aggregation was investigated. Microbiological analysis showed that 63% of the isolated LB were producing EPS, with the amount ranging from 8.21 to 43.13 mg/L. Chemical analysis of the extracts revealed the presence of polyphenols and flavonoids, more abundant in the hydromethanolic extract, which presented the highest content with 2.11 mg EGA/g of polyphenol and 1.64 mg EC/g of flavonoids and 1.71 mg EGA/g of polyphenol and 1.15 mg EC/g of flavonoids in the methanolic extract. Hydromethanolic extracts and EPS exhibited a more important activity than did the methanolic extract against EPEc. The combined action of EPS and extracts reduced the aggregation ability of EPEc and decreased the rate of their adhesion.

Caracterização da microbiota vaginal, intestinal e oral durante o período gestacional / Vaginal, gut and oral microbiota characterization during pregnancy

A simbiose desenvolvida entre seres vivos e microrganismos desempenha um importante papel na relação saúde-doença do hospedeiro. Neste sentido, o corpo humano abriga uma grande e diversa comunidade de microrganismos, sendo as mucosas vaginal, intestinal e oral as principais superfícies mucosas do corpo feminino que abrigam as comunidades bacterianas de fundamental importância para a mulher. Estes microrganismos atuam no desenvolvimento e modulação do sistema imune, na manutenção e otimização de vias metabólicas e competem por sítios de colonização, prevenindo que microrganismos patogênicos estabeleçam colonização. A composição da microbiota feminina varia com a idade, pH, secreção hormonal, ciclo menstrual, uso de anticoncepcional e atividade sexual. O presente estudo buscou caracterizar a composição da microbiota do corpo feminino durante o período gestacional, comparando os achados entre gestantes e não gestantes saudáveis, através de técnicas de biologia molecular. Foram selecionadas 60 mulheres saudáveis para o estudo e coletadas amostras de secreção vaginal, fezes e swab oral de cada participante. O DNA das amostras foi extraído e submetido à sequenciamento do gene 16S rRNA e quantificado através da técnica de PCR em tempo real. Das participantes selecionadas, 42 eram gestantes e 18 eram mulheres não gestantes em idade reprodutiva. Observamos que a quantificação total de bactérias na vagina não apresentou diferenças entre gestantes e não gestantes. Houve aumento na abundância de Lactobacillus no sítio vaginal, bactérias produtoras de butirato na microbiota intestinal e Streptococcus na microbiota oral de mulheres grávidas quando comparadas com mulheres não gestantes. Além disso, observamos que a composição e a disposição dos gêneros encontrados sofrem uma modificação, tal como aumento de gêneros relacionados com a manutenção da homeostase no grupo de mulheres gestantes. O período gestacional influencia positivamente na composição da microbiota, garantindo assim a prevalência de gêneros bacterianos responsáveis pela manutenção das condições ideais para o desenvolvimento da gestação saudável

The symbiosis developed between living organisms and microorganisms plays an important role in the health-disease relationship of the host. In this sense, the human body harbor a large and diverse community of microorganisms, the vaginal, intestinal and oral mucosa are the main mucosal surfaces of the female body that harbor bacterial communities of fundamental importance for women. These microorganisms act in the development and modulation of the immune system, in the maintenance and optimization of metabolic pathways and compete for colonization sites, preventing pathogenic microorganisms from establishing colonization. The composition of the female microbiota varies with age, pH, hormonal secretion, menstrual cycle, contraceptive use and sexual activity. The present study aimed to characterize the microbiota composition of the female body during the gestational period, comparing the findings between healthy and non - pregnant women through molecular biology techniques. Sixty healthy women were selected for the study and samples of vaginal secretion, stool and oral swab from each participant were collected. The DNA of the samples was extracted and submitted to the 16S rRNA gene sequencing and quantified by the real-time PCR technique. Were select, 42 were pregnant and 18 were non-pregnant women of reproductive age. We observed that the total quantification of bacteria in the vaginal samples did not present differences between pregnant and non-pregnant women. There was an increase in the abundance of Lactobacillus in the vaginal site, butyrate producing bacteria in the intestinal microbiota and Streptococcus in the oral microbiota of pregnant women when compared to nonpregnant women. In addition, we observed that the composition and arrangement of the genera found undergo a modification, such as an increase in genera related to the maintenance of homeostasis in the group of pregnant women. The pregnancy influences the composition of the microbiota, thus ensuring the prevalence of bacterial genera responsible for the maintenance of the ideal conditions for the development of healthy pregnancy

Use of probiotics in atopic dermatitis

SUMMARY Atopic dermatitis is a common skin disease. Its increased incidence has changed the focus of research on atopic dermatitis toward epidemiology, prevention, and treatment. Evidence suggests that intestinal microbiota plays an important role in the pathogenesis of atopic dermatitis inducing immunosuppression, but its exact mechanism is still unclear. Probiotics have been widely reported to act on the immune system. They are living microorganisms with immunomodulatory effects that stimulate Th1 cytokines and suppress the Th2 response, which are being researched for the treatment of several diseases. Probiotics most commonly used are part of the intestinal microflora like lactobacilli, bifidobacteria, and enterococci. We describe here a case of evident response to the use of probiotics in a girl with severe atopic dermatitis, with a significant change in severity scores of atopic dermatitis (BSA/SCORAD/FDLQI). Modulation of the intestinal microbiota with probiotics may offer a way to prevent or treat allergic diseases, including atopic dermatitis.

RESUMO A dermatite atópica é uma doença de pele comum. O aumento da incidência mudou o foco da pesquisa em dermatite atópica para epidemiología, prevenção e tratamento. Evidências sugerem que a microbiota intestinal desempenha um papel importante na patogênese da dermatite atópica, induzindo imunossupressão, mas o mecanismo exato ainda não está claro. Os probióticos foram amplamente divulgados para atuar no sistema imunológico. Eles são microrganismos vivos com efeitos imunomoduladores que estimulam as citocinas Th1 e suprimem a resposta Th2 que vem sendo pesquisada para o tratamento de diversas doenças. Probióticos mais comumente usados são parte da microflora intestinal como lactobacilos, bifidobactérias e enterococos. Descrevemos um caso de resposta evidente ao uso de probióticos em uma menina com dermatite atópica grave, com grande alteração nos escores de gravidade da dermatite atópica (BSA/Scorad/FDLQI). A modulação da microbiota intestinal com probióticos pode oferecer uma maneira de prevenir ou tratar doenças alérgicas, incluindo a dermatite atópica.

Identification of Vaginal Fluid Using Microbial Signatures / 法医学杂志

OBJECTIVES@#To investigate the specific microbial signatures in vaginal fluid.@*METHODS@#Vaginal fluid （16 samples）, saliva （16 samples）, feces （16 samples）, semen （8 samples）, peripheral blood （8 samples）, urine （5 samples）, and nasal secretion （4 samples） were collected respectively. The 16S rRNA genes of Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus jensenii, Lactobacillus iners, and Atopobium vaginae were amplified. PCR production was detected via a 3130xl Genetic Analyzer.@*RESULTS@#The detected number of Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus jensenii, Lactobacillus iners, and Atopobium vaginae were 15, 5, 8, 14, and 3 in all vaginal fluid samples, respectively. Lactobacillus crispatus and Lactobacillus jensenii existed specifically in vaginal fluid.@*CONCLUSIONS@#There is a potential application value to detect Lactobacillus crispatus and Lactobacillus jensenii for the identification of vaginal fluid.

Optimization of growth and bacteriocin production by Lactobacillus sakei subsp. sakei2a

Lactobacillus sakei subsp. sakei 2a is a bacteriocinogenic lactic acid bacterium isolated from Brazilian pork sausage, capable of inhibiting the growth of microbial pathogens, mainly Listeria monocytogenes. In order to optimize bacteriocin production for industrial applications, this study evaluated the effect of supplementation of MRS broth with glucose, Tween 20, Tween 80, sodium citrate, potassium chloride and cysteine, and effect of the initial pH and temperature of incubation of the medium on production of bacteriocins by L. sakei 2a. Adding glucose and Tween 20 to the medium, an initial pH of 5.0 or 5.5, and incubation temperatures of 25 °C or 30 °C resulted to the highest bacteriocin yields. Thus, a 2⁴ factorial design with the four variables was performed, and statistical analysis showed that it was an adequate model (R² = 0.8296). In the studied range, the four parameters significantly influenced bacteriocin production, with the maximum yield produced at an initial pH between 5.5 and 7.0, a temperature between 25 and 30 °C and supplementation of the MRS broth with glucose from 3.25 to 6.0 g L⁻¹ and Tween 20 from 0.575 to 1.15% (v/v). Response Surface Methodology analysis indicated that the highest bacteriocin production (12800 AU mL⁻¹) occurred in the MRS broth supplemented with 5.5 g L⁻¹ glucose and 1.05% Tween 20 at an initial pH of 6.28 and an incubation temperature of 25 °C. The amount of bacteriocin produced in commercial MRS broths under the same conditions was only 5600AU mL⁻¹.

Enumeration, identification and safety proprieties of lactic acid bacteria isolated from pork sausage / Enumeração, identificação e propriedades de segurança de bactérias ácido-lácticas isoladas de linguiça suína

Lactic Acid Bacteria (LAB) are indigenous microorganisms occurring in pork sausages. The utilization of selected autochthonous LAB may improve the safety of meat products. This study aims to enumerate and identify LAB in pork sausage and to characterize their safety properties, such as antimicrobial susceptibility and antibacterial activity. A total of 189 sealed packages of pork sausages were collected in seven municipalities (27 samples in each city) of Minas Gerais, Brazil. Microbiological analyses were performed to enumerate LAB. Two pre-selection criteria were applied to 567 isolates of LAB: catalase activity and tolerance to pH 2. A total of 32 strains of UFLA SAU were selected, characterized phenotypically and identified through 16S rDNA region sequencing. The susceptibility to antimicrobial and antibacterial activities of isolates was evaluated. The LAB count ranged from 3.079 to 8.987 log10 CFU/g. Lactobacillus plantarum and Lactobacillus paracasei were identified in the samples. UFLA SAU 11, 20, 34, 86, 131 and 258 showed a profile of susceptibility to four antimicrobials: erythromycin, ampicillin, chloramphenicol and gentamycin. In the antibacterial activity test, with exception of UFLA SAU 1, all other strains showed efficiency in inhibiting Escherichia coli, Salmonella Typhiand Listeria monocytogenes. In the statistical analysis there was interaction among strains of Lactobacillus against the pathogens tested. L. monocytogenes (P=0.05) was more sensitive to Lactobacillus strains and the highest inhibitory activity against this pathogen was achieved by strains UFLA SAU 135, 226, 238 and 258. Thus, UFLA SAU 11, 20, 34, 86, 131, 135, 226, 238 and 258 possess safety characteristics for application in meat products.

Bactérias ácido-lácticas (BAL) são microrganismos indígenas em linguiças. A utilização de selecionadas BAL autóctones pode melhorar a segurança dos produtos cárneos. Este estudo objetivou enumerar e identificar BAL em linguiças suínas e caracterizar suas propriedades de segurança, como a susceptibilidade antimicrobiana e a atividade antibacteriana. Um total de 189 embalagens fechadas de linguiça suína foi adquirido em sete municípios (27 amostras em cada cidade) de Minas Gerais, Brasil. Análises microbiológicas para a enumeração de BAL foram realizadas. Dois critérios de pré-seleção foram aplicados para os 567 isolados de BAL: atividade catalase e tolerância ao pH 2. Um total de 32 estirpes UFLA SAU foi selecionado, caracterizado fenotipicamente e identificado por meio do sequenciamento da região 16S rDNA. A susceptibilidade a antimicrobianos e a atividade antimicrobiana dos isolados foram avaliadas. Nas linguiças, a contagem de BAL variou de 3,079 a 8,987log10 UFC/g. Lactobacillus plantarum e Lactobacillus paracasei foram identificados nas amostras. UFLA SAU 11, 20, 34, 86, 131 e 258 apresentaram um perfil de suscetibilidade a quatro antimicrobianos: eritromicina, ampicilina, cloranfenicol e gentamicina. No teste de atividade antibacteriana, com exceção da UFLA SAU 1, todas as outras estirpes mostraram eficiência em inibir Escherichia coli, Salmonella Typhi e Listeria monocytogenes. Na análise estatística, houve interação entre estirpes de Lactobacillus contra os patógenos testados. L. monocytogenes (P=0,05) foi mais sensível às estirpes de Lactobacillus, e a maior atividade inibitória contra este patógeno foi apresentada por estirpes UFLA SAU 135, 226, 238 e 258. Assim, estirpes UFLA SAU 11, 20, 34, 86, 131, 135, 226, 238 e 258 possuem características de segurança para aplicação em produtos cárneos.

Identification and adhesion profile of Lactobacillus spp. strains isolated from poultry

In the aviculture industry, the use of Lactobacillus spp. as a probiotic has been shown to be frequent and satisfactory, both in improving bird production indexes and in protecting intestine against colonization by pathogenic bacteria. Adhesion is an important characteristic in selecting Lactobacillus probiotic strains since it impedes its immediate elimination to enable its beneficial action in the host. This study aimed to isolate, identify and characterize the in vitro and in vivo adhesion of Lactobacillus strains isolated from birds. The Lactobacillus spp. was identified by PCR and sequencing and the strains and its adhesion evaluated in vitro via BMM cell matrix and in vivo by inoculation in one-day-old birds. Duodenum, jejunum, ileum and cecum were collected one, four, 12 and 24 h after inoculation. The findings demonstrate greater adhesion of strains in the cecum and an important correlation between in vitro and in vivo results. It was concluded that BMM utilization represents an important technique for triage of Lactobacillus for subsequent in vivo evaluation, which was shown to be efficient in identifying bacterial adhesion to the enteric tract.

Adhesion, Autoaggregation and Hydrophobicity of Six Lactobacillus Strains.

Aims: The aim of this study was to assess probiotic attributes such as adhesion, auto aggregation, hydrophobicity and antibacterial activity of Lactobacillus strains from dairy products. Methodology: In this study, the autoaggregation, coaggregation, hydrophobicity and adhering abilities and antimicrobial activities of six Lactobacillus strains belonging to different species were assessed. Hydrophobicity was determined by bacterial adherence to hydrocarbons, xylene, n-hexadecane and chloroform. Results: The percentage of hydrophobicity of the strains range from 29.5% to 77.4% as measured by the described test. The autoaggregation among Lactobacillus strains range from 15.8% to 63.1%, while coaggregation range from 18.6% to 55.1%. Adhesion of the tested strains to buccal epithelial cells range from 8.0% to 50%. The tested Lactobacillus strains demonstrated variable inhibitory activity against pathogenic bacteria. Conclusion: Our findings indicated that one Lactobacillus strain expressed broad antibacterial activities against a group of bacterial pathogens and along 2 other strains exhibited ability to adhere to epithelial cells as shown by aggregation, coaggregation and hydrophobicity, indicating that such isolates can be good candidates for probiotic use.

Molecular characterization of lactobacilli isolated from fermented idli batter

Lactic acid bacteria are non pathogenic organism widely distributed in nature typically involved in a large number of spontaneous food fermentation. The purpose of this study was to characterize the bacteriocinogenic lactobacilli from fermented idli batter which can find application in biopreservation and biomedicine. Eight most promising lactobacilli were chosen from twenty two isolates based on their spectrum of activity against other lactic acid bacteria and pathogens. The eight lactobacilli were characterized based on the various classical phenotypic tests, physiological tests and biochemical tests including various carbohydrate utilization profiles. All isolates were homo fermentative, catalase, and gelatin negative. Molecular characterization was performed by RAPD, 16S rRNA analysis, 16S ARDRA, and Multiplex PCR for species identification. RAPD was carried out using the primer R2 and M13. Five different clusters were obtained based on RAPD indicating strain level variation. 16S rRNA analysis showed 99 to 100% homology towards Lactobacillus plantarum. The restriction digestion pattern was similar for all the isolates with the restriction enzyme AluI. The subspecies were identified by performing Multiplex PCR using species specific primer. Among the five clusters, three clusters were clearly identified as Lactobacillus plantarum subsp. plantarum, Lactobacillus pentosus, and Lactobacillus plantarum subsp. argentoratensis.

Identification of Lactobacillus plantarum, Lactobacillus pentosus and Lactobacillus fermentum from honey stomach of honeybee

This study aimed to isolate and identify Lactobacillus in the honey stomach of honeybee Apis dorsata. Samples of honeybee were collected from A. dorsata colonies in different bee trees and Lactobacillus bacteria isolated from honey stomachs. Ninety two isolates were Gram-stained and tested for catalase reaction. By using bacterial universal primers, the 16S rDNA gene from DNA of bacterial colonies amplified with polymerase chain reaction (PCR). Forty-nine bacterial 16S rDNA gene were sequenced and entrusted in GenBank. Phylogenetic analysis showed they were different phylotypes of Lactobacillus. Two of them were most closely relevant to the previously described species Lactobacillus plantarum. Other two phylotypes were identified to be closely related to Lactobacillus pentosus. However, only one phylotype was found to be distantly linked to the Lactobacillus fermentum. The outcomes of the present study indicated that L. plantarum, L. pentosus, and L. fermentum were the dominant lactobacilli in the honey stomach of honeybee A. dorsata collected during the dry season from Malaysia forest area - specifically "Melaleuca in Terengganu".

Effect of newly isolated Lactobacillus ingluviei ADK10, from chicken intestinal tract on acetaminophen induced oxidative stress in Wistar rats.

The total antioxidative activity of L. ingluviei ADK10 isolated from chicken intestine intact cells and cell free culture supernatant (CFCS) was 54- 67.95%. The ability to scavenge a,a-Diphenyl-b-Picrylhydrazyl free radical ranged from 71 and 64% in intact cells and CFCS respectively. Total reducing activity of bacteria was equivalent to 290 µM/L of cysteine. Reducing glutathione activity was equivalent to 93.95µg/mL. Oral administration of the strain at a dose of 109 cfu/kg body weight to acetaminophen induced oxidative stress in rats increased catalase, glutathione and superoxide dismutase activity in the blood, liver and kidney and lowered malondialdehyde level. The results indicate that L. ingluviei ADK10 has potential free radical scavenging activity for the treatment of oxidative stress related disease.

Identification of Lactobacillus spp. in colostrum from Chilean mothers

The biodiversity of Lactobacillus spp. in colostrum samples from 116 Chilean mothers was analyzed by PCR and 16S rDNA sequencing. Lactobacilli were isolated in 55.3% of the samples, with concentrations of 3.33 ± 0.55 (log CFU/ml). The predominant species were L. plantarum (64%), L. fermentum (16%) and L. pentosus (9%). 28% of the isolated strains were resistant to gastric pH and bile salts, suggesting that they could be used as probiotics.

Se analizó la biodiversidad de especies de Lactobacillus en muestras de calostro de 116 madres chilenas mediante PCR y secuenciación del rDNA 16S. Se aislaron lactobacilos en 55,3% de las muestras, con concentraciones de 3,33 ± 0,55 (log UFC / ml). Las especies predominantes fueron L. plantarum (64%), L. fermentum (16%) y L. pentosus (9%). 28% de las cepas aisladas fueron resistentes a pH gástrico y a las sales biliares, lo que sugiere que podrían ser utilizados como probióticos.

Identification and in vitro production of Lactobacillus antagonists from women with or without bacterial vaginosis

Lactobacilli isolated from the vaginal tract of women with and without bacterial vaginosis (BV) were identified and characterized for the production of antagonists. Bacterial samples were isolated from healthy women (N = 16), from patients with clinical complaints but without BV (N = 30), and from patients with BV (N = 32). Identification was performed using amplified ribosomal DNA restriction analysis. Production of antagonistic compounds was evaluated by the double-layer diffusion technique using Gram-positive (N = 9) and Gram-negative bacteria (N = 6) as well as yeast (N = 5) as indicator strains. Of a total of 147 isolates, 133 were identified as pertaining to the genus Lactobacillus. Lactobacillus crispatus was the species most frequently recovered, followed by L. johnsonii and L. jensenii. Statistical analysis showed that L. crispatus was more frequent in individuals without BV (P < 0.05). A higher production of antagonists was noted in L. crispatus isolates from healthy women (P < 0.05). More acidic local pH and higher H2O2 production by isolated lactobacilli from healthy women suggest these mechanisms as the possible cause of this antagonism. In conclusion, a significant correlation was detected between the presence and antagonistic properties of certain species of Lactobacillus and the clinical status of the patients.

Spectrum of Lactobacillus species present in healthy vagina of Indian women.

Background & objectives: Lactobacilli are depleted in vagina of women suffering from recurring episodes of bacterial vaginosis with vaginal pH >5. With the objective of making available probiotic lactobacilli for replenishment in such women, a study was undertaken to isolate and characterize the Lactobacilli present in women with eco-healthy vagina in Delhi. No information is so far available on the species of Lactobacilli resident in vagina of women in India. Methods: Vaginal swabs were taken from 80 women with informed consent after ethical approval and grown in MRS broth. Gram-positive, catalase-negative bacilli generating about 200 bp amplicon by PCR with Lactobacillus genus specifi c primers were further characterized by employing species specifi c primers followed by sequencing of 16S rDNA. Isolates of the same species were differentiated by random amplifi ed polymorphic DNA (RAPD) profi les. Results: The predominant species isolated were L. reuteri present in 26 (32.5%) women, L. fermentum in 20 (25%), and L. salivarius in 13 (16.25%) women. Sequencing of 16S rDNA of 20 isolates showed that except for two isolates of L. plantarum, sequences of the remaining agreed well with PCR identifi cation. None of the isolates had similar RAPD profi le. Interpretation & conclusions: Our fi ndings showed lactobacilli species present in healthy vagina of women in India differ from those reported from other countries. This information would be useful to development of probiotic tablets seeking to replenish the missing lactobacilli for reproductive health of women in India.

Prevalência e caracterização de espécies de lactobacilos vaginais em mulheres em idade reprodutiva sem vulvovaginites / Prevalence and characterization of vaginal lactobacillus species in women at reproductive age without vulvovaginitis

OBJETIVO: identificar espécies de lactobacilos isolados do conteúdo vaginal de mulheres saudáveis e assintomáticas; determinar as espécies mais prevalentes e caracterizá-las fenotipicamente. MÉTODOS: lactobacilos foram isolados em meio seletivo a partir de amostras de conteúdo vaginal de 135 mulheres, sem queixa de corrimento e com diagnóstico laboratorial negativo para infecções vaginais, acompanhadas em um ambulatório de Planejamento Familiar. Os isolados foram identificados por PCR multiplex e, quando necessário, submetidos ao sequenciamento do gene RNAr 16S. Foram também avaliados quanto à acidificação do meio de cultura, à produção de ácido láctico, de H2O2, bacteriocinas e a capacidade de adesão às células epiteliais. RESULTADOS: oitenta e três cepas de lactobacilos foram isoladas e identificadas, sendo as espécies predominantes L. crispatus (30,1%), L. jensenii (26,5%), L. gasseri (22,9%) e L. vaginalis (8,4%). Apenas 20 destes isolados não produziram H2O2 em quantidades detectáveis. Das 37 linhagens selecionadas para teste de adesão a células epiteliais, 12 apresentaram adesão entre 50 a 69%, 10 apresentaram 70% ou mais, e as restantes pouca ou nenhuma adesão. Nenhum dos isolados produziu bacteriocinas. CONCLUSÕES: as espécies de lactobacilos mais prevalentes em mulheres sem vulvovaginites, isoladas em meio de cultura seletivo e identificadas por métodos moleculares, foram L. crispatus, L. jensenii e L. gasseri. Além de mais frequentes, tais linhagens também apresentaram melhor produção de H2O2 e atingiram menores valores de pH em meio de cultura.

PURPOSE: to identify species of lactobacillus isolated from the vaginal contents of healthy and asymptomatic women, determining the most prevalent species and characterizing them phenotypically. METHODS: lactobacillus have been isolated in selective milieu from samples of the vaginal contents of 135 women without complaints of vaginal secretion, and with negative laboratorial diagnosis of vaginal infection, followed up at an outpatient clinic. After being identified by multiplex PCR, the isolates have been submitted to RNAr 16S gene sequencing, when necessary. They have also been evaluated concerning the production of lactic acid, H2O2, bacteriocins and the ability to adhere to epithelial cells. RESULTS: eight-three lactobacillus strains were isolated and identified, L. crispatus (30.1%), L. jensenii (26.5%), L. gasseri (22.9%) e L. vaginalis (8.4%), being the prevalent species. Only 20 of those isolates did not present H2O2 production, in detectable amounts. From the 37 strains selected for the test of adhesion to the epithelial cells, 12 presented 50 to 69% of adhesion, 10 presented 70% or more, and the remaining, little or no adhesion at all. None of the tested strains produced bacteriocins. CONCLUSIONS: the lactobacillus species more prevalent in women without vulvovaginitis, isolated in selective culture milieu and identified by molecular methods were L. crispatus, L. jensenii and L. gasseri. Besides the fact of being more prevalent, these strains also presented better production of H2O2, and reached lower pH values in the culture milieu.

Prevalencia e caracterização de especies de lactobacilos vaginais em mulheres saudaveis em idade reprodutiva / Prevalence and characterization of lactobacilos vaginais species in healthy women at reproductive age

A microflora vaginal de mulheres saudáveis em idade reprodutiva é composta por uma variedade de bactérias aeróbias e anaeróbias, mas as espécies dominantes são os lactobacilos (bacilos de Dõderlein), que exercem significante influência sobre a microbiota local. Além de restringir o crescimento de patógenos competindo pelo espaço e nutrientes, os lactobacilos produzem substâncias antimicrobianas como ácidos orgânicos, peróxido de hidrogênio (H2O2) e bacteriocinas. Esta atividade antagonista é importante na proteção contra várias infecções, principalmente a vaginose bacteriana (VB). Objetivos: Identificar as espécies de lactobacilos isolados do conteúdo vaginal de mulheres saudáveis e assintomáticas e determinar as espécies mais prevalentes e caracterizá-las quanto à produção de ácido láctico, peróxido de hidrogênio (H2O2) e sua capacidade de adesão às células do epitélio vaginal. Métodos: Foram isoladas 83 linhagens de lactobacilos de amostras de conteúdo vaginal de 135 mulheres, sem queixa de corrimento e com diagnóstico laboratorial negativo para infecções vaginais, acompanhadas no ambulatório de Planejamento Familiar da Faculdade de Ciências Médicas - UNICAMP. As linhagens isoladas foram identificadas por PCR multiplex e, quando necessário, submetidas ao sequenciamento do gene RNAr 16S. Foram então avaliadas quanto à produção de ácido láctico, de H2O2, bacteriocinas e a capacidade de adesão às células epiteliais. Resultados: A espécie predominante foi L. crispatus presente em 30,1% das mulheres, seguida de L. jensenii (26,5%), L. gasseri (22,9%) e L. vaginalis (8,4%)...

The vaginal microflora of healthy women is composed of a large variety of aerobic and anaerobic bacteria. The dominant species is a group of lactobacilli (Doderlein's bacillus), which has a significant effect on vaginal microbiota, curtailing the growth of pathogens competing for space and nutrients. The lactobacilli specie produces various antimicrobial substances that include organic acids, hydrogen peroxide (H202) and bacteriocins. Objectives: Identify the prevalence of the different species of lactobacilli isolated from the vagina of healthy asymptomatic women, determine the most prevalent species and characterize them regarding the production of lactic acid, hydrogen peroxide (H2O2) and their capacity to adhere to vaginal epithelial cells. Methods: Eightythree strains of lactobacilli were isolated from the vagina of 135 women, with no complaints of vaginal discharge and negative laboratory diagnosis for vaginal infection, who were being followed up at the Family Planning clinic of the Medical school, Unicamp. The isolates were identified using multiplex polymerase chain reaction (PCR) and, when...

Aislamiento y estudio de diferentes especies de lactobacillus de la cavidad oral / Isolation and study of different species of oral lactobacilli

El estudio de los gérmenes del género Lactobacillus de la cavidad bucal ha sido principalmente orientado al desarrollo e interpretación de tests destinados a la cuantificación de los mismos. Consideramos de interés profundizar en el conocimiento de las características de la colonización de estos microorganismos con la puesta a punto de las técnicas de identificació a nivel de especie. En una segunda instancia se planteó la posibilidad de correlacionar los morfotipos coloniales obtenidos habitualmente en el medio de Rogosa y los resultados de la especiación. Se estudiaron 120 muestras de saliva, con técnicas de cultivo en superficie y cuantificación. Se aislaron Lactobacillus en 102 muestras y de todas ellas se realizó estudio de los morfotipos y especiación de los mismos. Lactobacillus casei, Lactobacillus acidophilus (en ese orden), fueron las especies aisladas con mayor frecuencia. Es posible, trabajando en condiciones estandarizadas, establecer una correlación entre los diversos morfotipos que se observan en el medio de Rogosa y las especies de aislamiento más frecuente. Esto permite manejar información adicional en los resultados de los tests predictivos de recuento en placa de Lactobacillus